Poly(ADP-ribose) quantification at the femtomole level in mammalian cells

Anal Biochem. 1995 Jul 1;228(2):245-51. doi: 10.1006/abio.1995.1346.

Abstract

ADP-ribose polymers were isolated from living mammalian cells, separated by polyacrylamide gel electrophoresis, visualized in the gel with a novel silver staining agent, and quantified by computer-aided scanning densitometry. This method detects as little as approximately 40 fmol of ADP-ribose polymers of a particular size class and reduces the gel exposure times required for conventionally radiolabeled polymers from 2 months to about 1 h. The method also detects polymers with slow turnover which may be underestimated by techniques requiring metabolic radiolabeling of poly(ADP-ribose).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Diphosphate Ribose / metabolism*
  • Biopolymers
  • Cell Line
  • Glycoside Hydrolases
  • Humans
  • Microchemistry
  • Reference Standards
  • Sensitivity and Specificity
  • Silver Staining / methods*

Substances

  • Biopolymers
  • Adenosine Diphosphate Ribose
  • Glycoside Hydrolases
  • poly ADP-ribose glycohydrolase